title: X-ray scattering experiments with high-flux X-ray source coupled rapid mixing microchannel device and their potential for high-flux neutron scattering investigations.
abstract: Small-angle X-ray scattering provides global, shape-sensitive structural information about macromolecules in solution. Its extension to time dimension in the form of time-resolved SAXS investigations and combination with other time-resolved biophysical methods contributes immensely to the study of protein dynamics. TR-SAXS can also provide unique information about the global structures of transient intermediates during protein dynamics. An experimental set-up with low protein consumption is essential for an extensive use of TR-SAXS experiments on protein dynamics. In this direction, a newly developed 20-microchannel microfluidic continuous-flow mixer was combined with SAXS. With this set-up, we demonstrate ubiquitin unfolding dynamics after rapid mixing with the chaotropic agent Guanidinium-HCl within milliseconds using only ∼ 40 nanoliters of the protein sample per scattering image. It is suggested that, in the future, this new TR-SAXS platform will help to increase the use of time-resolved small-angle X-ray scattering, wide-angle X-ray scattering and neutron scattering experiments for studying protein dynamics in the early millisecond regime. The potential research field for this set-up includes protein folding, protein misfolding, aggregation in amyloidogenic diseases, function of intrinsically disordered proteins and various protein-ligand interactions.
authors.affiliation: Structural Dynamics of (Bio)chemical Systems, MPI-BPC, Am Fassberg 11, 37077, Goettingen, Germany.
authors.forename: R
authors.initials: R
authors.lastname: Jain
authors.forename: M
authors.initials: M
authors.lastname: Petri
authors.forename: S
authors.initials: S
authors.lastname: Kirschbaum
authors.forename: H
authors.initials: H
authors.lastname: Feindt
authors.forename: S
authors.initials: S
authors.lastname: Steltenkamp
authors.forename: S
authors.initials: S
authors.lastname: Sonnenkalb
authors.forename: S
authors.initials: S
authors.lastname: Becker
authors.forename: C
authors.initials: C
authors.lastname: Griesinger
authors.forename: A
authors.initials: A
authors.lastname: Menzel
authors.forename: T P
authors.initials: TP
authors.lastname: Burg
authors.forename: S
authors.initials: S
authors.lastname: Techert
mesh_terms: D033363:Neutron Diffraction; D011506:Proteins / Q000737:chemistry*; D053838:Scattering, Small Angle; D014961:X-Ray Diffraction
chemical_list: D011506:Proteins